miR-296对人肺腺癌A549细胞恶性生物学行为的影响

Effect of miR-296 on the malignant behavior of lung adenocarcinoma cells

目的探讨微小RNA-296(miR-296)对人肺腺癌A549细胞增殖、凋亡和迁移能力的影响及其潜在的分子机制。方法将miR-296表达载体、抑制剂及相应阴性对照分别转染人肺腺癌A549细胞,实时PCR检测miR-296在不同细胞中的表达情况,CCK8法和平板克隆实验检测miR-296对A549细胞增殖能力的影响,流式细胞术检测细胞凋亡率,Transwell法检测细胞迁移能力,Western blotting检测S100A4在各组A549细胞中的表达情况。结果与相应的对照组比较,转染了miR-296表达载体的A549细胞miR-296表达显著上调,细胞增殖和迁移能力明显下降,凋亡率明显上升,S100A4蛋白表达下调,差异均有统计学意义(均P<0.05);而转染了miR-296抑制剂的A549细胞miR-296表达显著下调,细胞增殖和迁移能力明显增强,凋亡率明显下降,S100A4蛋白表达上调,差异均有统计学意义(均P<0.05)。结论miR-296通过靶向调控S100A4表达来抑制A549细胞增殖、迁移,促进凋亡。

Objective To investigate the effect of microRNA-296(miR-296)on the proliferation,apoptosis,and migration potential of lung adenocarcinoma cells and its underlying molecular mechanisms.Methods miR-296 overexpressing vector,miR-296 inhibitor,and corresponding negative control,were transfected into human lung adenocarcinoma A549 cells.miR-296 expression was assessed by realtime polymerase chain reaction(RT-PCR).Cell proliferation was analyzed by the CCK8 cell proliferation assay kit and clone formation.Apoptosis and migration were evaluated by flow cytometry and Transwell assay,respectively,and the expression of S100A4 was assessed by Western blotting.Results Compared with the corresponding negative control group,the proliferative and migratory potential of A549 was dramatically inhibited,the apoptosis rate was significantly elevated,and S100A4 levels were significantly decreased in miR-296 overexpressing cells(all P<0.05).In contrast,A549 proliferation and migration were dramatically enhanced,cell apoptosis was significantly decreased,and the expression of S100A4 was significantly increased upon miR-296 inhibition(all P<0.05).Conclusion miR-296 inhibits the proliferation and migration,and promotes apoptosis of A549 cells,probably via the regulation of S100A4 expression.